Matthew Wilson
Last active: 3/24/2020

CRISPR/Cas9 engineering of albino cystinuria Type A mice.

Beckermann TM, Welch RC, Williams FM, Mortlock DP, Sha F, Ikizler TA, Woodard LE, Wilson MH
Genesis. 2020 58 (5): e23357

PMID: 32078250 · PMCID: PMC7781090 · DOI:10.1002/dvg.23357

Cystinuria Type A is a relatively common genetic kidney disease occurring in 1 in 7,000 people worldwide that results from mutation of the cystine transporter rBAT encoded by Slc3a1. We used CRISPR/Cas9 technology to engineer cystinuria Type A mice via genome editing of the C57BL/6NHsd background. These mice are an improvement on currently available models as they are on a coisogenic genetic background and have a single defined mutation. In order to use albinism to track Cas9 activity, we co-injected gRNAs targeting Slc3a1 and tyrosinase (Tyr) with Cas9 expressing plasmid DNA into mouse embryos. Two different Slc3a1 mutational alleles were derived, with homozygous mice of both demonstrating elevated urinary cystine levels, cystine crystals, and bladder stones. We used whole genome sequencing to evaluate for potential off-target editing. No off-target indels were observed for the top 10 predicted off-targets for Slc3a1 or Tyr. Therefore, we used CRISPR/Cas9 to generate coisogenic albino cystinuria Type A mice that could be used for in vivo imaging, further study, or developing new treatments of cystinuria.

© 2020 Wiley Periodicals, Inc.

MeSH Terms (10)

Amino Acid Transport Systems, Basic Amino Acid Transport Systems, Neutral Animals CRISPR-Cas Systems Cysteine Cystinuria Disease Models, Animal Mice Mice, Inbred C57BL Mutation

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